Skardal Biofabrication Lab
Research Overview
Our research program focuses on the design and implementation of extracellular matrix-inspired hydrogel biomaterials for the biofabrication of tissue and tumor organoids, organ-on-a-chip systems, and cancer-on-a-chip systems for drug screening, disease modeling, and personalized medicine. Our team’s work has broad applicability across tissue types and diseases but has focused primarily in development of cancer models and recently neural models. Our biomaterial systems are largely hyaluronic acid-based with additional custom ECM component addons, synthetically modified in the lab. These are often combined with microfluidic devices for fluid handling capabilities. In particular, we have a significant interest in patient biospecimen-derived models that can facilitate and improve current precision medicine and precision oncology efforts in the clinic.
Research Projects
Glioblastoma subtype evolution in patient-derived glioblastoma tumor organoids
Glioblastoma (GBM) is a lethal, incurable form of cancer in the brain that universally recurs more aggressively even with maximally aggressive surgery followed by chemoradiotherapy. These tumors are extremely heterogenous with regions of genetically distinct subclones that evolve differently over time and in response to treatments making designing effective therapies for each individual patient difficult. Here are using GBL cell lines and a patient-specific ex vivo tumor-on-a-chip system to analyze tumor heterogeneity and drift over time to predict clonal evolution for patients, which could subsequently have substantial impact on treatment decisions.
Personalized drug screening in patient-derived tumor organoids
Our team has developed a technology portfolio comprised of a range of bioengineered 3D tissue and tumor models, which in the last several years has expanded to include tumor organoids created from patient tumor biospecimens. These patient-derived tumor organoids (PTOs) fill a critical experimental gap, facilitating screening studies that can provide patient-specific empirical data to better predict a patient’s drug response, and that addresses the heterogeneity between patients and individual tumors. We combine organ micro-engineering with microfluidics, and PTOs formed to generate tumor-on-a-chip (TOC) systems. To date, the lab has created PTOs from a range of tumor types, including colorectal, appendiceal, lung, melanoma, myeloma, glioma, ovarian, and mesothelioma tumors.
Immune-enhanced organoids for immunotherapy screening in vitro
Reconstructing the patient’s own tumor in the form of patient derived tumor organoids (PTOs) recapitulates the tumor microenvironment by incorporating tumor cells along with associated stroma and tumor infiltrating leucocytes (TILs). Due to the variable infiltration of tumors by (TILs) with inconsistent functional status, PTOs were thought as not being suitable to recapitulate the complex interactions between tumors and the patient’s own immune system. To address this limitation, we have conceived of mixing lymph node-derived cells and tumor-derived cells from the same patient creating personalized tumor/node organoids, allowing for individual patient tumor and stroma and immune system to remain viable and operational, recapitulating the interaction between host patient and its own tumor. Importantly, these immune-enhanced organoids allow for successful screening of immunotherapy agent such as immune checkpoint blockade therapies.
Metastasis-on-a-chip
Metastatic disease remains one of the primary reasons for cancer-related deaths, yet the majority of in vitro cancer models focus on the primary tumor sites. Our lab has developed a series of metastasis-on-a-chip devices that house multiple bioengineered three-dimensional (3D) organoids, established by a 3D photopatterning technique employing extracellular matrix-derived hydrogel biomaterials. Specifically, cancer cells begin in primary site organoid, which resides in a single microfluidic chamber connected to multiple downstream chambers in which other target site organoids such as liver, lung, endothelial constructs, or other tissue types are housed. Under recirculating fluid flow, tumor cells grow in the primary site, eventually enter circulation, and can be tracked via fluorescent imaging. Studies on this platform can be implemented to better understand the mechanisms underlying metastasis, perhaps resulting in the identification of targets for intervention.
Neurovascular unit (NVU)-on-a-chip
Our group is working on establishing a more advanced NVU within a microfluidic device, complete with a functional BBB. These models will allow for the direct study of BBB mass transport, effects of fluid flow, shear stress and pressure, and systemic versus direct-to-brain drug delivery in both normal and diseased states.
Extracellular matrix bioinks for 3D bioprinting
3D bioprinting has advanced rapidly since its inception, particularly with respect to hardware platforms, yet much less attention has been paid to understanding the interactions between the biomaterials, or bioinks, with the hardware and the cells beyond simple viability metrics. Unfortunately, most researchers employ outdated biomaterials that were never designed to be compatible with the dynamic events encountered during bioprinting. As a result, there is a need for novel biomaterials with more advanced, stimuli-responsive mechanical properties that will be simple to implement in a variety of bioprinter platforms, thus enabling acceleration of technologies that will drive biomanufacturing of replacement tissue products for patients.
Active Funding:
1R21CA229027 - NCI IMAT Program
XCELL Biologix
The Ohio State University Comprehensive Cancer Center
Completed/Past Funding:
Grant Number: W81XWH-15-9-001
Source: s/RegenMed Development Organization (ReMDO) via MTEC
Title: Development of a Universal Bioink with Tunable Mechanical Properties for Regenerative Medicine Additive Manufacturing of Clinical Products
Role: PI
Performance Period: 11/04/2016 – 10/30/2021
Annual Direct Costs: $637,486
Time Commitment: 1.8 calendar months (15% effort)
Goal: To develop and engineer a modular system allowing for customization of bioink biomaterials for bioprinting that can be deployed in all common bioprinting hardware modalities and be tailored to create and support the majority of the tissue types in the human body – both for regenerative applications and organoid technologies for diagnostics, personalized medicine, and drug development.
Grant Number: n/a
Source: Wake Forest Breast Cancer Center of Excellence
Title: An Immuno-Organoid Platform for ex vivo Testing of Novel Immunotherapies for TNBC Patients
Role: PI
Performance Period: 1/01/2019 – 12/31/2019
Annual Direct Costs: $100,000
Time Commitment: 0.6 calendar months (5% effort)
Goal: Utilize parallel murine models and murine organoid triple negative breast cancer models to direct immune-enhancing of patient-derived breast cancer tumor organoids and subsequent techniques to reduce immune-suppression in triple negative breast cancer, enabling immune checkpoint blockade therapies.
Grant Number: n/a
Source: Myeloma Crowd Foundation
Title: Rapid and Personalized Prediction of Myeloma Response to Chemotherapy Using CD Organoids
Role: Co-PI
Performance Period: 1/01/2019 – 12/31/2019 (option to renew)
Annual Direct Costs: $150,000
Time Commitment: 1.2 calendar months (10% effort)
Goal: Develop supportive microenvironment conditions to support difficult to maintain ex vivo myeloma cell populations. Deploy organoids created in personalized chemosensitivity screening studies and correlate with patient outcomes.
Grant Number: n/a
Source: Wake Forest Clinical and Translational Science Institute
Title: A tunable thixotropic hydrogel bioink for bioprinting of functional tissue analogs
Performance Period: 4/01/2018 – 3/31/2019
Role: PI
Annual Direct Costs: $40,000
Time Commitment: no salary
Goal: We propose to employ, query, and quantify the contributions of inter-polymer/protein forces (hydrogen bonding versus covalent bonding) in generating thixotropic hydrogel bioinks, subsequently assessing how these forces influence bioprinting parameters during tissue construct biofabrication. We will characterize and optimize contributions of non-covalent, hydrogen bond-based interactions within thixotropic extracellular matrix (ECM)-based hydrogel bioinks. Subsequently we will employ and validate bioinks by bioprinting viable and functional tissue constructs (liver construct and heart patch).
Grant Number: n/a
Source: Comprehensive Cancer Center at Wake Forest Baptist
Title: Patient tumor-derived tumor-on-a-chip technology for determining metastatic potential and response to chemotherapy prior to initiation of treatment
Role: PI
Performance Period: 5/01/2017 – 11/30/2019
Annual Direct Costs: $10,000
Time Commitment: no salary
Goal: Employ genetic screening to identify drugable biomarkers in patient gastrointestinal tumor biopsies. Use cells from these biopsies to create 3D tumor organoids with which to test biomarker-driven drugs for efficacy for each patient case.
Grant Number: 2017-614-001
Source: Medical Technology Enterprise Consortium (MTEC)
Title: Pre-clinical Assessment of Bioprinted Human Skin for Wound Healing and Skin Regeneration
Role: Co-I
Performance Period: 11/28/2017 – 11/19/2018
Annual Direct Costs: $193,548
Time Commitment: 0.84 calendar months (7% effort)
Goal: The overall goal of the project is to bioprint full-thickness human skin with hair follicle appendages, microvasculature, immune cells and pigmentation and use it as a skin graft in immunodeficient rats.
Grant Number: 1 R33 CA202822-01
Source: NIH
Title: Bioengineered Lung Tumor Organoids for Development of Personalized Medicine
Role: Co-I
Performance Period: 04/12/2016 – 03/31/2019
Annual Direct Costs: $248,983
Time Commitment: 0.96 calendar months (8% effort)
Goal: Personalized oncology, whereby tumor DNA is sequenced to identify actionable gene mutations, is poised to become a standard process in cancer treatment,but is dependent on the availability of sufficient amounts of intact tumor cell DNA. We propose to bioprint lung organoids that will recapitulate the in vivo lung microenvironment in order to successfully expand a small number of freshly isolated lung cancer cells in vitro.
Grant Number: W81XWH-13-2-0054
Source: DOD USAMRAA
Title: Amniotic Fluid-Derived Stem Cells for Enhanced Wound Healing
Role: Co-I
Performance Period: 09/23/2013 – 09/22/2019 (NCE)
Annual Direct Costs: $0
Time Commitment: 2.64 calendar months (22% effort)
Goal: The goal of this work is to develop a method that will permit the use of allogeneic source of fetal stem cells, and novel hydrogels for clinical management of burn wounds, allowing wound healing treatment to achieve fast and comprehensive wound coverage that results in functional and cosmetically superior skin.
Grant Number: 1 R21 CA28933-01A1
Source: NIH
Title: Development of a CF10 Predictive Gene Signature in CRC Organoids
Role: Co-I
Performance Period: 02/02/2018 – 01/31/2020
Annual Direct Costs: $130,000
Time Commitment: 0.96 calendar months (8% effort)
Goal: This project investigates whether a new polymeric fluoropyrimidine, CF10, is effective for treating colorectal cancer (CRC) that is non-responsive to 5-fluorouracil (5-FU). Our preliminary studies show CF10 is much more potent than 5-FU and is effective in models of CRC sub-types that are non-responsive to 5-FU. We will systematically evaluate response of CRC cells and patient tumors using novel tumor organoid technology to establish in what CRC sub-types CF10 treatment provides a therapeutic advantage.
Title: INtegrated Organoid Testing System, (INGOTS)
Role: Co-I
Supporting Agency: Defense Threat Reduction Agency/SPAWAR
Performance Period: 03/28/2013 – 10/01/2018
Level of Funding (total costs): $17,750,269
Goals: INGOTS will be comprised of four interconnected microscale bioreactors, each containing fully-functional, three dimensional (3D) human tissue constructs (organoids). INGOTS will allow for the application of test agents at the individual organoid or whole body system level and will employ both commercially available human cells and blood substitutes.
Title: Patient-specific 3D tumor organoids for glioblastoma multiforme precision medicine
Role: Principal Investigator
Supporting Agency: Wake Forest Brain Tumor Center of Excellence
Performance Period: 05/01/2016 – 06/30/2017
Level of Funding (total costs): $30,000
Goals: The goal is to demonstrate and implement a GBM organoid system within the framework of clinical precision medicine, by demonstrating biomarker- and mutation-based drug targeting in 3D patient-derived GBM tumor models. To accomplish this goal, patient-derived GBM organoids will be fabricated, and genetic profiles will be used to inform customized drug screening. These customized GBM treatments will be assessed for responsiveness using quantitative analysis of tumor growth, reduction, and migration in 3D space, and viability/apoptosis.
Title: Patient-specific Bioengineered Lung Tumor Organoids to support personalized medicine
Role: Co-Investigator
Supporting Agency: Center for Public Health Genomics
Performance Period: 08/01/2015 – 08/01/2016
Level of Funding (total costs): $25,000
Goal: To employ bioprinted lung organoids to support growth of non-small cell lung cancer biopsies in order to increase cellular yield that can be used for genetic screens for precision medicine.
Title: A Three-Dimensional Liver Microtumor Organoid Platform for Anti-Cancer Drug Development
Role: Principal Investigator
Supporting Agency: Golfers Against Cancer
Performance Period: 03/01/2013 – 03/01/2015
Level of Funding (total costs): $40,000
Goals: In a rotating bioreactor, liver organoids will be created, inside of which reside metastatic colon carcinoma cells. In this 3-D environment, cancer cells behave as they would when in the body, providing a superior human cell-based in vitro testing platform for screening potential drug candidates in comparison to traditional 2-D cultures or animals.
Title: CTSI Translational Pilot - A patient-specific tumor-on-a-chip platform for screening precision medicine-driven therapies
Role: Principal Investigator
Supporting Agency: Wake Forest Baptist Medical Center – Clinical and Translational Science Institute
Performance Period: 02/01/2016 – 04/01/2017
Level of Funding (total costs): $40,000
Goals: To demonstrate the utility of using tumor models created using cells from actual patient tumor biopsies to screen drug therapies for a given patient, thereby identifying the most effective treatment. Single colorectal cancer tumor organoids are created within colon constructs and screened using drugs commonly employed against colorectal cancer.
Title: WFIRM – Promoting Innovative Discoveries – Intramural Pilot Funding Program
Role: Principal Investigator
Supporting Agency: Wake Forest Institute for Regenerative Medicine
Performance Period: 10/01/2014 – 10/01/2015
Level of Funding (total costs): $25,000
Goals: To merge microfluidic device technology with photo-patterned hydrogel biomaterials to create a high-throughput system of 3-D tumor and tissue organoids for drug testing and metastasis exploration. Initial pilot work focuses on colon carcinoma metastases in liver organoids. Post-pilot work will expand to other tissue/tumor types and the biological mechanisms that play important roles in cancer that can potential therapeutic targets for intervention.
